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表面活性蛋白D高表达对脂多糖诱导人肾小管上皮细胞单核细胞趋化蛋白1表达的影响
  • ISSN号:1001-7097
  • 期刊名称:《中华肾脏病杂志》
  • 时间:0
  • 分类:R739.41[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]武汉大学人民医院肾内科,430060
  • 相关基金:国家自然科学基金(30670985)
中文摘要:

目的 研究表面活性蛋白D(SP-D)高表达对脂多糖诱导人肾小管上皮细胞(HK-2)单核细胞趋化蛋白1(MCP-1)表达的影响及其机制.方法 间接免疫荧光、Western印迹及RT-PCR检测SP-D蛋白及mRNA在HK-2细胞内的表达.脂多糖在不同浓度(0、0.1、1、2、5、10 mg/L)作用8 h及5 mg/L脂多糖作用HK-2细胞不同时间(0、2、4、8、16、24 h),采用Western印迹和实时定量PCR检测SP-D表达,ELISA和实时定量PCR检测MCP-1表达.脂质体转染法将pEE14-hSP-D质粒转染HK-2细胞,筛选稳定转染细胞株,Westem印迹检测转染后SP-D蛋白表达.采用5 mg/L脂多糖刺激pEE14-hSP-D稳定转染的HK-2细胞8 h,ELISA和实时定量PCR检测HK-2细胞MCP-1表达.结果 HK-2细胞表达SP-D.脂多糖可诱导MCP-1 mRNA及蛋白表达显著增高(P〈0.05);并町诱导SP-D mRNA及蛋白表达显著降低(P〈0.05).转染pEEl4-hSP-D质粒使HK-2细胞高表达SP-D,并可下调脂多糖诱导的MCP-1表达(P〈0.01).结论 SP-D可通过下调脂多糖诱导的HK-2细胞MCP-1的表达,从而在肾脏炎性反应中起重要作用.

英文摘要:

Objective To investigate the effect of surfactant protein D(SP-D)overexpression on lipopolysaccharide(LPS)-induced monocyte chemoattractant protein-1(MCP-1)expression in human renal proximal tubular epithelial cells(HK-2)and its mechanism. Methods HK-2 cells were treated with LPS at various concentrations (0, 0.1, 1, 2, 5, 10 mg/L)for 8 h and at 5 mg/L for various time points(0, 2, 4, 8, 16, 24 h). Expression of SP-D was detected by Western blotting and real-time PCR. Expression of MCP-1 was determined by ELISA and real-time PCR. Human SP-D cDNA eukaryotic expression vector pEE14-hSP-D was transfected to HK-2 cells. The changes in transfected cells of SP-D protein were observed by Western blotting. Expression of MCP-1 was detected by ELJSA and real-time PCR. Results SP-D was expressed in HK-2 cells. The levels of SP-D protein and mRNA in HK-2 cells were significantly decreased after treatment with LPS(P〈0.05). Expression of MCP-1 protein and mRNA was increased remarkably after treatment with LPS(P〈0.05). HK-2 cells transfected with pEE14-hSP-D showed up-regulated expression of SP-D. The overexpression of SP-D inhibited the LPS-inducedexpression of MCP-1(P〈0.01). Conclusions SP-D inhibits LPS-induced expression of MCP-1 in HK-2 cells. SP-D may play an important role in the modulation of renal inflammation.

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期刊信息
  • 《中华肾脏病杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中华医学会
  • 主编:
  • 地址:广州市中山二路74号
  • 邮编:510089
  • 邮箱:cmaszb@mail.sysu.edu.cn
  • 电话:020-87331532
  • 国际标准刊号:ISSN:1001-7097
  • 国内统一刊号:ISSN:44-1217/R
  • 邮发代号:46-106
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:26419